LOCUS       sequence-249721-        3234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes VPR activation domain as a Type 3b part to be used in the 
            MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3234)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 3234)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK3b_021
FEATURES             Location/Qualifiers
     source          1..3234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      131..719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     620..639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             834..983
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             990..1010
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1830..2399
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     terminator      complement(2419..2453)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2524..3183)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3098..3117
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga tcaatcatga
       61 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      241 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      781 tctttcctgc gttatcccct gattctgtgg ataaccgtag tcggtctcat tctgatgctt
      841 tagacgattt tgacttagat atgcttggtt cagacgcgtt agacgacttc gacctagaca
      901 tgttaggctc agatgcattg gacgacttcg atttagatat gttgggctcc gatgccctag
      961 atgactttga tctagatatg ctaggtagtc ccaaaaagaa gaggaaagtg ggatcccagt
     1021 atctgcccga cacagatgat agacaccgaa tcgaagagaa acgcaagcga acgtatgaaa
     1081 ccttcaaatc gatcatgaag aaatcgccct tctcgggtcc gaccgatccc aggcccccac
     1141 cgagaaggat tgcggtcccg tcccgctcgt cggccagcgt gccgaagcct gcgccgcagc
     1201 cctacccctt cacgtcgagc ctgagcacaa tcaattatga cgagttcccg acgatggtgt
     1261 tcccctcggg acaaatctca caagcctcgg cgctcgcacc agcgcctccc caagtccttc
     1321 cgcaagcgcc tgccccagcg cctgcaccgg caatggtgtc cgccctcgca caggcccctg
     1381 cgcccgtccc cgtgctcgcg cctggaccgc cccaggcggt cgctccaccg gctccgaagc
     1441 cgacgcaggc cggagaggga acactctccg aagcacttct tcaactccag tttgatgacg
     1501 aggatcttgg agcactcctt ggaaactcga cagaccctgc ggtgtttacc gacctcgcgt
     1561 cagtagataa ctccgaattt cagcagcttt tgaaccaggg tatcccggtc gcgccacata
     1621 caacggagcc catgttgatg gaataccccg aagcaatcac gagacttgtg acgggagcgc
     1681 agcggcctcc cgatcccgca cccgcacctt tgggggcacc tggcctccct aacggacttt
     1741 tgagcggcga cgaggatttc tcctccatcg ccgatatgga tttctcagtc ttgctgtcac
     1801 agatttccag cggctctggc agcggcagcc gggattccag ggaagggatg tttttgccga
     1861 agcctgaggc cggctccgct attagtgacg tgtttgaggg ccgcgaggtg tgccagccaa
     1921 aacgaatccg gccatttcat cctccaggaa gtccatgggc caaccgccca ctccccgcca
     1981 gcctcgcacc aacaccaacc ggtccagtac atgagccagt cgggtcactg accccggcac
     2041 cagtccctca gccactggat ccagcgcccg cagtgactcc cgaggccagt cacctgttgg
     2101 aggatcccga tgaagaaacg agccaggctg tcaaagccct tcgggagatg gccgatactg
     2161 tgattcccca gaaggaagag gctgcaatct gtggccaaat ggacctttcc catccgcccc
     2221 caaggggcca tctggatgag ctgacaacca cacttgagtc catgaccgag gatctgaacc
     2281 tggactcacc cctgaccccg gaattgaacg agattctgga taccttcctg aacgacgagt
     2341 gcctcttgca tgccatgcat atcagcacag gactgtccat cttcgacaca tctctgtttg
     2401 gatcctgaga ccagaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     2461 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc
     2521 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     2581 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     2641 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     2701 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaaa cgaaaaacat
     2761 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     2821 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     2881 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     2941 ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat
     3001 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     3061 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     3121 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     3181 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccgg
//
